2,2-Bis(hydroxymethyl)-2,2',2''-nitrilotriethanol
[CAS# 6976-37-0]

Identification

• Classification: Pharmaceutical intermediate >> Heterocyclic compound intermediate >> Pyrimidine compound >> Alcohol
• Name: 2,2-Bis(hydroxymethyl)-2,2',2''-nitrilotriethanol
• Synonyms: Bis(2-hydroxyethyl)iminotris(hydroxymethyl)methane; BIS-TRIS
• Molecular Formula: C₈H₁₉NO₅
• Molecular Weight: 209.24
• Protein Sequence: G
• CAS Registry Number: 6976-37-0
• EC Number: 230-237-7
• SMILES: C(CO)N(CCO)C(CO)(CO)CO

Properties

• Melting point: 103-104 °C
• Solubility: 2000 mM (water)

Safety Data

• Hazard Symbols: GHS05;GHS07 Danger
• Risk Statements: H315-H318-H319-H335
• Safety Statements: P261-P264-P264+P265-P271-P280-P302+P352-P304+P340-P305+P351+P338-P305+P354+P338-P317-P319-P321-P332+P317-P337+P317-P362+P364-P403+P233-P405-P501

Hazard Classification

Hazard	Class	Category Code	Hazard Statement
Eye irritation	Eye Irrit.	2	H319
Skin irritation	Skin Irrit.	2	H315
Specific target organ toxicity - single exposure	STOT SE	3	H335
Serious eye damage	Eye Dam.	1	H318
Substances or mixtures corrosive to metals	Met. Corr.	1	H290
Specific target organ toxicity - single exposure	STOT SE	3	H336

Discovery and Applications

2,2-Bis(hydroxymethyl)-2,2',2''-nitrile triethanol, also known as BIS-TRIS, is an important zwitterionic buffer. It was developed as part of an effort to create a variety of effective biological buffers that met the criteria established by Norman E. Good in the 1960s. Good's buffers were known for their minimal interaction with biological systems, chemical stability, and nontoxicity, making them ideal for a variety of biochemical and biological applications. BIS-TRIS has a stable buffering capacity designed to meet the need for effective control of pH within the physiological range.

The molecular formula of BIS-TRIS is C8H19NO5 and its molecular weight is 209.24 g/mol. Its structure features a tris(hydroxymethyl)aminomethane backbone with additional bis(hydroxymethyl) groups. This structure gives BIS-TRIS excellent buffering properties, making it effective over a pH range of 5.8 to 7.2 and a pKa value of approximately 6.5 at 25°C. This pH range is particularly useful for many biochemical applications, ensuring a stable environment for a variety of reactions and processes.

BIS-TRIS is widely used in electrophoresis buffers to help maintain a consistent pH during protein and nucleic acid separations. Its stable buffering capacity ensures accurate and reproducible results, which is critical for techniques such as SDS-PAGE and agarose gel electrophoresis. Using BIS-TRIS in electrophoresis buffers can improve the resolution and separation of biomolecules.

In cell culture applications, BIS-TRIS helps maintain the pH of the culture medium within the physiological range, promoting optimal cell growth and viability. Its non-toxic properties ensure that it does not interfere with cellular processes, making it a reliable buffer for maintaining consistent experimental conditions in cell-based studies. The stability of BIS-TRIS in cell culture media helps reproducibility of experimental results.

BIS-TRIS is used in a variety of biochemical assays to stabilize pH, which is critical for the activity and stability of enzymes and other biomolecules. Its buffering capacity ensures that assay conditions remain optimal, allowing for accurate and reliable measurements of biochemical activity. The non-interfering properties of BIS-TRIS make it a key ingredient in enzyme research and other biochemical analyses.

During protein purification, maintaining a stable pH is essential to preserve protein structure and function. BIS-TRIS is used in chromatography techniques and other purification protocols to buffer the system and improve the yield and purity of isolated proteins. Its compatibility with different purification methods makes it a versatile tool in protein chemistry.

In structural biology, BIS-TRIS is used in techniques such as X-ray crystallography and NMR spectroscopy to study the structure of proteins and other biomolecules. Its stability and low ionic strength make it an ideal buffer for obtaining high-quality structural data without interfering with the biomacromolecules being analyzed.

The high buffering capacity allows BIS-TRIS to effectively maintain a stable pH, ensuring consistent experimental results. With an appropriate pH range, BIS-TRIS can be used in a variety of experimental conditions, making it a versatile buffer. BIS-TRIS is generally non-toxic to cells and does not interfere with biochemical reactions, making it suitable for sensitive biological applications.

References

2023. Complexes of Cu(II), Co(II), and Zn(II) Terephthalates with Hydroxyalkylamines. Russian Journal of Coordination Chemistry, 49(8).
DOI: 10.1134/s1070328423600389